纳米粒子介导的BMP-2基因转染对移植血管内膜增生的抑制作用  

Inhibitory effect of BMP-2 gene transfection mediated by nanoparticles on proliferation of graft vascular inner membrane

作　　者：李智[1] 郎晓讴[1] 宦大为[2] 孙虓[2] 

机构地区：[1]沈阳医学院奉天医院普外科,辽宁沈阳110024 [2]沈阳医学院奉天医院病理科,辽宁沈阳110024

出　　处：《吉林大学学报（医学版）》2013年第1期82-86,197,共5页Journal of Jilin University：Medicine Edition

基　　金：沈阳医学院青年人才科技基金资助课题(2010)

摘　　要：目的:研究人骨形态发生蛋白2(BMP-2)基因局部转染后,BMP-2基因蛋白过表达对血管平滑肌细胞(VSMC)增殖的抑制作用,探讨防治移植静脉再狭窄的新途径。方法:以包载BMP-2基因的聚乳酸-羟基乙酸共聚物(PLGA)的纳米级粒子混合物(BMP-2-PLGA)转染培养的兔VSMC,作为BMP-2-PLGA纳米粒子组,同时设空白PLGA纳米粒子组及对照组,MTT法测定细胞增殖活性,流式细胞术分析细胞周期。建立自体静脉移植兔模型,随机分成转基因组、空白载体粒子组及单纯静脉移植对照组,分别于术后3、7、14和28d取出移植静脉,Verhoeff染色检测血管内膜厚度,Western blotting法检测BMP-2蛋白的表达,免疫组织化学法检测BMP-2及增殖细胞核抗原(PCNA)表达。结果:与对照组比较,BMP-2-PLGA纳米粒子组细胞发生明显的G1期阻滞(P<0.05),细胞增殖抑制率及细胞凋亡率增加(P<0.05)。与空白载体粒子组及单纯静脉移植对照组比较,转基因组兔血管内膜平均厚度明显减少(P<0.05),空白载体粒子组与单纯静脉移植对照组间比较差异无统计学意义(P>0.05)。与空白载体粒子组及单纯静脉移植对照组比较,转基因组兔移植静脉组织BMP-2蛋白表达于术后3、7和14d明显增多(P<0.05),PCNA表达于术后7～28d明显降低(P<0.01);空白粒子组与单纯静脉移植对照组比较差异无统计学意义(P>0.05)。结论:BMP-2基因的表达能够有效抑制自体静脉移植后内膜增生及VSMC的增殖,促进体外培养VSMC的凋亡。Objective To investigate the inhibitory effect of bone morphogenetic protein 2 （BMP-2） gene transfection mediated by nanoparticles （NP） on proliferation of vascular smooth muscle cells （VSMC） in rat vein grafting model, and to provide a new method to prevent restenosis. Methods The VSMC were transduced in vitro with NP BMP-2 DNA complex prepared with PLGA and used as BMP-2-PLGA group, meanwhile simple PLGA group and control group were set up. The cell proliferation was determined by flow cytometry. Autogenous vein graft models were established in 72 rabbits by transplanting internal branch of jugular vein to carotid artery, then divided into BMP-2 group, empty vector group and simple graft control group. The grafted veins were harvested at 3, 7, 14 and 28 d respectively after operation. The thickness of vascular inner membrane was detected with Verhoeff staining. The exogenous BMP-2 protein expression in veins was determined by Western blotting. The expressions of proliferating cell nuclear antigen （PCNA） and BMP-2 were detected by immunohistochemistry. Results Compared with control group, the inhibitory rate of cell proliferation and apoptotic rate in BMP-2 PLGA group were increased obviously （P〈0.05） and the cell cycle was arrested in G1 phase （P〈0.05）. Compared with empty vector group and simple graft control group, the thickness of vascular inner membrane in BMP-2 group was decreased （P〈0.01）, but there was no significant difference between empty vector group and simple graft control group. Compared with empty vector group and simple graft control group, the BMP-2 protein expressions in rabbit vein graft tissue in BMP-2 group 3, 7, 14 d after operation were significantly increased （P〈0.05）; the PCNA expressions were significantly decreased 7--28 d after operation （P〈0.05） ; but there was no significant difference between empty vector group and simple graft control group. Conclusion Expression of BMP-2 gene can prevent intimal hyperplasia （IH） and VSMC prolifer

关 键 词：骨形态发生蛋白2基因 纳米粒子 移植 

分 类 号：R654.3[医药卫生—外科学]